The present disclosure relates to an information processing apparatus, an information processing method, a program, and a method of correcting the intensity of a fluorescence spectrum.
To measure the characteristics of microparticles such as cells, an apparatus (for example, a flow cytometer) that irradiates a laser beam to microparticles marked by a fluorescent pigment and measures the intensity or a pattern of fluorescence coming from the excited fluorescent pigment is used. As a technology for analyzing the characteristics of microparticles in further detail, a technology, called multi-color measurement, of marking microparticles using a plurality of fluorescent pigments and measuring fluorescence coming from each fluorescent pigment radiated with a laser beam using optical detectors with different light reception wavelength bands has also been used. Further, a transparent wavelength band of an optical filter provided in each optical detector to restrict the light reception wavelength band is designed in accordance with the fluorescent wavelength of the fluorescence coming from the fluorescent pigment to be measured.
For example, fluorescein isothiocyanate (FITC) or phycoerythrin (PE) is used as the fluorescent pigment. When a fluorescence spectrum obtained by irradiating a laser beam to microparticles marked with such a fluorescent pigment is measured, the presence of the fluorescence wavelength bands overlapping one another is confirmed. That is, even when the fluorescence obtained by irradiating the laser beam to the microparticles is separated in accordance with wavelength bands by an optical filter in the multi-color measurement, a fluorescent component coming from a fluorescent pigment other than a target fluorescent component is considered to leak in the fluorescence spectrum detected by each optical detector. When the leakage of the fluorescence occurs, a deviation may occur between a fluorescence intensity measured by each optical detector and the fluorescence intensity of the fluorescence actually coming from the target fluorescent pigment. As a result, a measurement error occurs.
To correct such a measurement error, a fluorescence correction process (compensation) of subtracting the fluorescence intensity corresponding to the leakage of the fluorescence from the fluorescence intensity measured by the optical detector is performed. The fluorescence correction process is a process of performing correction (hereinafter referred to as fluorescence correction) on the measured fluorescence intensity such that the fluorescence intensity measured by the optical detector approximates the fluorescence intensity of the fluorescence actually coming from the target fluorescent pigment. For example, as a method of performing the fluorescence correction, Japanese Unexamined Patent Application Publication No. 2003-83894 discloses a method of mathematically correcting a fluorescence intensity.
According to the method disclosed in Japanese Unexamined Patent Application Publication No. 2003-83894, the fluorescence intensity of the fluorescence actually coming from the target fluorescent pigment is calculated using a vector which has a fluorescence intensity (detection value) measured by each optical detector as a component and applying an inverse matrix of a correction matrix set in advance in the vector. The correction matrix is also called a leakage matrix. The correction matrix is a matrix which is prepared by analyzing a fluorescence wavelength distribution of microparticles singly marked with respective fluorescent pigments and arranging the fluorescence wavelength distributions of each fluorescent pigments as a row vector.